The feasibility of single molecule Millikan Sequencing for de novo sequencing of mammalian genomes for under $1,000 will be evaluated. This novel sequencing-by-synthesis approach measures the increased charge as nucleotides are added to a single DNA template attached to a tethered bead without any need for labels. Opposing electrical, hydrodynamic and restoring forces result in bead motion that is a function of the length of DNA attached to the bead. Simultaneous optical detection of an array of millions of beads undergoing chain elongation will allow high-throughput sequencing. Model calculations and preliminary results indicate that this method should enable accurate, long read length and label-free DNA sequencing. The lack of labels leads to negligible reagent costs while the relatively simple dark-field optics leads to a low-cost instrument. Long read lengths will result in low genome assembly cost. The single template approach eliminates the need for emulsion or bridge PCR, making sample preparation much easier and cheaper. The aim of the proposed two-year exploratory project is to demonstrate the feasibility of sequencing a single DNA template by monitoring changes in the intrinsic charge of a growing DNA chain. PUBLIC HEALTH RELEVANCE: Nucleic acid sequence information is critical to medical research and to basic biological studies. The goal of this project is to demonstrate the feasibility of DNA sequencing system based on a label-free detection approach, single molecule Millikan Sequencing, which may be capable of de novo sequencing of mammalian genomes for under $1,000. Such a platform will allow researchers and clinicians to perform the translational research ultimately required to improve human health.